Extended Data Fig. 2: AMPK inactivation renders cells sensitive to ferroptotic cell death.
From: Energy-stress-mediated AMPK activation inhibits ferroptosis
a, Western blot showing the levels of ACC (S79) and AMPK (T172) phosphorylation in ACHN cells treated with erastin for 24 h or cultured in cystine-free media for 36 h with and without compound C (10 μM). b, Cell death measurement in AMPK WT and DKO ACHN cell lines treated with 100 nM RSL3 for 16 h. c, d, Cell death measurement in RCC4 cells treated with 10 μM of compound C, cell death inhibitors, and 5 μM of erastin for 24 h (c) or in cystine free media for 36 h (d). Ferr-1: 1 μM ferrostatin-1; Z-V: 20 μM Z-VAD-FMK; Nec: 2 μM necrostatin-1; NAC: 5 mM N-acetyl cysteine. e, Western blot showing the AMPK expression in RCC4 cell as indicated. f–h, Cell death measurement in AMPK WT and DKO RCC4 cell lines treated with 5 μM of erastin for 24 h (f), cultured in cystine free media for 24 h (g), or treated with 100 nM of RSL3 for 24 h (h). i–k, Lipid peroxidation measurement in AMPK WT and DKO RCC4 cells treated with 5 μM of erastin for 18 h (i), cultured in cystine free media for 18 h (j), or treated with 100 nM of RSL3 for 12 h (k). l, Cell death measurement in indicated RCC4 cells cultured with cystine free media for 24 h. Data show the mean ± s.d., n = 3 independent experiments. Statistical analysis was performed using unpaired, two-tailed t-test. Numerical source data are provided in Source Data Extended Data Fig. 2. Scanned images of unprocessed blots are shown in Source Data Extended Data Fig. 2.
